Substituted piperidines as renin inhibitors

ABSTRACT

The present invention relates to compounds of the general formula (I) and the salts thereof, preferably the pharmaceutically acceptable salts thereof; in which R has the meaning explained in the description, a process for their preparation and the use of these compounds as medicines, especially as renin inhibitors.

FIELD OF THE INVENTION

The present invention relates to novel substituted 4-phenyl piperidines,to processes for their preparation and to the use of the compounds asmedicines, in particular as renin inhibitors.

BACKGROUND OF THE INVENTION

Piperidine derivatives for use as medicines are known, for example fromWO97/09311. However, especially with regard to renin inhibition, thereis still a need for highly potent active ingredients. In this context,the improvement of a compound's pharmacokinetic properties, resulting inbetter oral bioavailability, and/or it's overall safety profile are atthe forefront. Properties directed towards better bioavailability are,for example, increased absorption, metabolic stability or solubility, oroptimized lipophilicity. Properties directed towards a better safetyprofile are, for example, increased selectivity against drugmetabolizing enzymes such as the cytochrome P450 enzymes.

DETAILED DESCRIPTION OF THE INVENTION

The invention therefore provides substituted 4-phenyl piperidines of thegeneral formula

and their salts, preferably their pharmaceutically acceptable salts, inwhich

R is

C₂₋₈-alkenyl,C₂₋₈-alkynyl,C₀₋₈-alkyl-carbonyl-optionally N-mono-C₁₋₈-alkylated amino-C₁₋₈-alkyl,C₃₋₈-cycloalkyl-C₀₋₈-alkyl,C₁₋₈-alkyl-sulfonyl-C₁₋₈-alkyl,optionally N-mono- or N,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₈-alkyl,optionally O—C₁₋₈-alkylated carboxyl-C₀₋₈-alkyl,optionally N and/or N′ mono-, di- or tri-C₁₋₈-alkylatedureido-C₁₋₈-alkyl,heterocyclylcarbonyl-C₀₋₈-alkyl orheterocyclyl-C₀₋₈-alkyl;each of said radicals may be substituted, preferably by 1-4 substituentsindependently selected fromC₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl,cyano,halogen,hydroxyl,oxo,trifluoromethoxy andtrifluoromethyl.

The meaning of “C₀-alkyl” in the above (and hereinafter) mentionedC₀₋₈-alkyl groups is a bond or, if located at a terminal position, ahydrogen atom.

Examples of C₁₋₈-alkyl radicals, which may be linear or branched, aremethyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, pentyl and hexyl respectively. Examples of C₁₋₈-alkoxyradicals, which may be linear or branched, are radicals such as methoxy,ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy andtert-butoxy. Examples of C₂₋₈-alkenyl radicals, which may be linear orbranched, are, for example, vinyl and propenyl. Examples of C₂₋₈-alkynylradicals, which may be linear or branched, are, for example, ethynyl.Examples of O—C₁₋₈-alkylated carboxyl are radicals such asmethoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, iso propoxycarbonyl,n-butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl andtert-butoxycarbonyl. Examples of C₀₋₈-alkylcarbonylamino are for exampleformylamino, acetylamino, n-propionylamino, isopropionylamino,n-butylcarbonylamino, isobutylcarbonylamino, sec-butylcarbonylamino andtert-butylcarbonylamino. Examples of optionally N and/or N′ mono-, -di-or -tri-C₁₋₈-alkylated ureido are radicals such as ureido,1-methyl-ureido, 3-methyl-ureido, trimethyl-ureido, 1-ethyl-ureido,3-ethyl-ureido, triethyl-ureido, 1-ethyl-3-methyl-ureido,3-ethyl-1-methyl-ureido. Examples of optionally N-mono- orN,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₈-alkyl are radicals such ascarbamoyl, N-methyl carbamoyl, N-ethyl carbamoyl, N-propyl carbamoyl,N,N-di-methyl carbamoyl, N,N-di-ethyl carbamoyl, N,N-di-propylcarbamoyl, carbamoyl-methyl, N-methyl carbamoyl-methyl, N-ethylcarbamoyl-methyl, N-propyl carbamoyl-methyl, N,N-di-methylcarbamoyl-methyl, N,N-di-ethyl carbamoyl-methyl, N,N-di-propylcarbamoyl-methyl, carbamoyl-ethyl, N-methyl carbamoyl-ethyl, N-ethylcarbamoyl-ethyl, N-propyl carbamoyl-ethyl, N,N-di-methylcarbamoyl-ethyl, N,N-di-ethyl carbamoyl-ethyl, N,N-di-propylcarbamoyl-ethyl, carbamoyl-2-propyl, N-methyl carbamoyl-2-propyl,N-ethyl carbamoyl-2-propyl, N-propyl carbamoyl-2-propyl, N,N-di-methylcarbamoyl-2-propyl, N,N-di-ethyl carbamoyl-2-propyl, N,N-di-propylcarbamoyl-2-propyl, carbamoyl-(2-methyl-2-butyl), N-methylcarbamoyl-(2-methyl-2-butyl), N-ethyl carbamoyl-(2-methyl-2-butyl),N-propyl carbamoyl-(2-methyl-2-butyl), N,N-di-methylcarbamoyl-(2-methyl-2-butyl), N,N-di-ethyl carbamoyl-(2-methyl-2-butyl),N,N-di-propyl carbamoyl-(2-methyl-2-butyl).

The term heterocyclyl refers to 3-8 membered monocyclic, saturated orunsaturated heterocyclic radicals having 1 to 4 nitrogen and/or 1 or 2sulfur or oxygen atoms, for example saturated or unsaturatedN-containing C₃₋₈-heterocyclyl. The heterocyclyl radicals may besubstituted one or more times, such as, for example, substituted once ortwice by C₁₋₈-alkoxy, C₁₋₈-alkyl, cyano, halogen, hydroxy,trifluoromethoxy or trifluoromethyl. Heterocyclyl radicals whichcomprise a nitrogen atom may be linked either via the N atom or via a Catom to the remainder of the molecule. Examples of such heterocyclylradicals are imidazolyl, morpholinyl, oxetanyl, oxiranyl, pyrazolyl,pyridyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrazolyl,thiazolyl and triazolyl. Examples of preferred heterocyclic radicals aremorpholinyl, tetrazolyl and triazolyl.

Halogen is fluoro, chloro, bromo or iodo.

Cycloalkyl refers to a saturated cyclic hydrocarbon radicals having 3 to8 carbon atoms, for example cyclopropyl, cyclobutyl or cyclopentyl andmay be unsubstituted or substituted once or twice by C₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkyl, optionally halogen substituted C₁₋₈-alkyl orhalogen.

Salts are primarily the pharmaceutically usable or nontoxic salts ofcompounds of formula (I). The term “pharmaceutically useable salts”encompasses salts with inorganic or organic acids, such as hydrochloricacid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid,citric acid, formic acid, maleic acid, acetic acid, succinic acid,tartaric acid, methanesulfonic acid, p-toluenesulfonic acid and thelike.

Salts of compounds having salt-forming groups are in particular acidaddition salts, salts with bases, or, in the presence of a plurality ofsalt-forming groups, in some cases also mixed salts or internal salts.

Such salts are formed, for example, from compounds of formula (I) withan acidic group, for example a carboxyl or sulfonyl group, and are, forexample, the salts thereof with suitable bases such as non-toxic metalsalts derived from metals of group Ia, Ib, IIa and IIb of the PeriodicTable of the Elements, for example alkali metal, in particular lithium,sodium, or potassium, salts, alkaline earth metal salts, for examplemagnesium or calcium salts, and also zinc salts and ammonium salts,including those salts which are formed with organic amines, such asoptionally hydroxy-substituted mono-, di- or trialkylamines, inparticular mono-, di- or tri(lower alkyl)amines, or with quaternaryammonium bases, e.g. methyl-, ethyl-, diethyl- or triethylamine, mono-,bis- or tris(2-hydroxy(lower alkyl))amines, such as ethanol-, diethanol-or triethanolamine, tris(hydroxymethyl)methylamine or2-hydroxy-tert-butylamine, N,N-di(lower alkyl)-N-(hydroxy(loweralkyl))amine, such as N,N-di-N-dimethyl-N-(2-hydroxyethyl)amine, orN-methyl-D-glucamine, or quaternary ammonium hydroxides such astetrabutyl ammoniumhydroxide. The compounds of formula (I) having abasic group, for example an amino group, may form acid addition salts,for example with suitable inorganic acids, e.g. hydrohalic acid such ashydrochloric acid, hydrobromic acid, sulfuric acid with replacement ofone or both protons, phosphoric acid with replacement of one or moreprotons, e.g. orthophosphoric acid or metaphosphoric acid, orpyrophosphoric acid with replacement of one or more protons, or withorganic carboxylic, sulfonic or phosphonic acids or N-substitutedsulfamic acids, e.g. acetic acid, propionic acid, glycolic acid,succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid,fumaric acid, malic acid, tartaric acid, gluconic acid, glucaric acid,glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelicacid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid,2-acetoxybenzoic acid, embonic acid, nicotinic acid, isonicotinic acid,and also amino acids, for example the alpha-amino acids mentioned above,and also methanesulfonic acid, ethanesulfonic acid,2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid,benzenesulfonic acid, 4-methylbenzenesulfonic acid,naphthalene-2-sulfonic acid, 2- or 3-phosphoglycerate, glucose6-phosphate, N-cyclohexylsulfamic acid (with formation of thecyclamates) or with other acidic organic compounds such as ascorbicacid. Compounds of formula (I) having acidic and basic groups may alsoform internal salts.

Salts obtained may be converted to other salts in a manner known per se,acid addition salts, for example, by treating with a suitable metal saltsuch as a sodium, barium or silver salt, of another acid in a suitablesolvent in which an inorganic salt which forms is insoluble and thusseparates out of the reaction equilibrium, and base salts by release ofthe free acid and salt reformation.

The compounds of formula (I), including their salts, may also beobtained in the form of hydrates or include the solvent used for thecrystallization.

For the isolation and purification, pharmaceutically unsuitable saltsmay also find use.

The compounds of formula (I) also include those compounds in which oneor more atoms are replaced by their stable, non-radioactive isotopes;for example a hydrogen atom by deuterium.

The compounds of the formula (I) also include compounds that have beennitrosated through one or more sites such as oxygen (hydroxylcondensation), sulfur (sulfydryl condensation) and/or nitrogen. Thenitrosated compounds of the present invention can be prepared usingconventional methods known to one skilled in the art. For example, knownmethods for nitrosating compounds are described in WO2004/098538 A2.

The compounds of the formula (I) also include compounds that have beenconverted at one or more sites such that a nitrate-ester-containinglinker is attached to an existing oxygen and/or nitrogen. Such“nitroderivatives” of the compounds of the present invention can beprepared using conventional methods known to one skilled in the art. Forexample, known methods for converting compounds into theirnitroderivatives are described in WO2007/045551 A2.

The compounds of formula (I) have at least four asymmetric carbon atomsand may therefore be in the form of optically pure diastereomers,diastereomeric mixtures, diastereomeric racemates, mixtures ofdiastereomeric racemates or as meso compounds. The invention encompassesall of these forms. Diastereomeric mixtures, diastereomeric racemates ormixtures of diastereomeric racemates may be separated by customaryprocedures, for example by column chromatography, thin-layerchromatography, HPLC and the like.

The compounds of formula (I) may also be prepared in optically pureform. The separation into antipodes can be effected by procedures knownper se, either preferably at an earlier synthetic stage by saltformation with an optically active acid, for example (+)- or(−)-mandelic acid and separation of the diastereomeric salts byfractional crystallization, or preferably at a relatively late stage byderivatizing with a chiral auxiliary building block, for example (+)- or(−)-camphanoyl chloride, and separation of the diastereomeric productsby chromatography and/or crystallization and subsequent cleavage of thebonds to give the chiral auxiliary. The pure diastereomeric salts andderivatives may be analysed to determine the absolute configuration ofthe piperidine present with common spectroscopic procedures, and X-rayspectroscopy on single crystals constitutes a particularly suitableprocedure.

It is possible for the configuration at individual chiral centres in acompound of formula (I) to be inverted selectively. For example, theconfiguration of asymmetric carbon atoms which bear nucleophilicsubstituents, such as amino or hydroxyl, may be inverted by second-ordernucleophilic substitution, if appropriate after conversion of the bondednucleophilic substituent to a suitable nucleofugic leaving group andreaction with a reagent which introduces the original substituents, orthe configuration at carbon atoms having hydroxyl groups can be invertedby oxidation and reduction, analogously to the process in the Europeanpatent application EP-A-0 236 734. Also advantageous is the reactivefunctional modification of the hydroxyl group and subsequent replacementthereof by hydroxyl with inversion of configuration.

The compound groups mentioned below are not to be regarded as closed,but rather parts of these compound groups may be exchanged with oneanother or with the definitions given above or omitted in a sensiblemanner, for example to replace general by more specific definitions. Thedefinitions are valid in accordance with general chemical principles,such as, for example, the common valences for atoms.

The compounds of formulae (I), (IA) and (IB) can be prepared in ananalogous manner to preparation processes disclosed in the literature.Similar preparation processes are described for example in WO 97/09311and in WO06/103275. Details of the specific preparation variants can befound in the examples.

Preferred compounds are those of the general formulae (IA) or (IB)

and the salts thereof, preferably the pharmaceutically acceptable saltsthereof, are compounds in which R is as defined above for the compoundsof the formula (I).

A further, preferred group of compounds of the formula (I), or morepreferably of the formula (IA) or (IB), and the salts thereof,preferably the pharmaceutically acceptable salts thereof, are compoundsin which

R is

C₀₋₈-alkyl-carbonyl-optionally N-mono-C₁₋₈-alkylated amino-C₁₋₈-alkyl,optionally substituted, preferably by 1-2 substituents independentlyselected fromC₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl,cyano,halogen,hydroxyl,oxo,trifluoromethoxy andtrifluoromethyl.

A further, preferred group of compounds of the formula (I), or morepreferably of the formula (IA) or (IB), and the salts thereof,preferably the pharmaceutically acceptable salts thereof, are compoundsin which

R is

optionally N-mono- or N,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₈-alkyl,optionally substituted, preferably by 1-2 substituents independentlyselected fromC₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl,cyano,halogen,hydroxyl,oxo,trifluoromethoxy andtrifluoromethyl.

A further, preferred group of compounds of the formula (I), or morepreferably of the formula (IA) or (IB), and the salts thereof,preferably the pharmaceutically acceptable salts thereof, are compoundsin which

R is

optionally N and/or N′ mono-, di- or tri-C₁₋₈-alkylatedureido-C₁₋₈-alkyl,optionally substituted, preferably by 1-2 substituents independentlyselected fromC₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl,cyano,halogen,hydroxyl,oxo,trifluoromethoxy andtrifluoromethyl.

A further, preferred group of compounds of the formula (I), or morepreferably of the formula (IA) or (IB), and the salts thereof,preferably the pharmaceutically acceptable salts thereof, are compoundsin which

R is

heterocyclyl-C₀₋₈-alkyl,optionally substituted, preferably by 1-2 substituents independentlyselected fromC₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl,cyano,halogen,hydroxyl,oxo,trifluoromethoxy andtrifluoromethyl.R is particularly preferablyC₀₋₈-alkyl-carbonyl-optionally N-mono-C₁₋₈-alkylated amino-C₁₋₈-alkyl,optionally N-mono- or N,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₈-alkyl orheterocyclyl-C₀₋₈-alkyl.R is very particularly preferablyC₀₋₄-alkyl-carbonyl-optionally N-mono-C₁₋₄-alkylated amino-C₁₋₄-alkyl,optionally N-mono- or N,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₄-alkyl,morpholinyl-C₀₋₄-alkyl, tetrazolyl-C₀₋₄-alkyl or triazolyl-C₀₋₄-alkyl.

Prodrug derivatives of the compounds described herein are derivativesthereof which on in vivo use liberate the original compound by achemical or physiological process. A prodrug may for example beconverted into the original compound when a physiological pH is reachedor by enzymatic conversion. Possible examples of prodrug derivatives areesters of freely available carboxylic acids, S- and O-acyl derivativesof thiols, alcohols or phenols, the acyl group being defined as herein.Preferred derivatives are pharmaceutically acceptable ester derivativeswhich are converted by solvolysis in physiological medium into theoriginal carboxylic acid, such as, for example, lower alkyl esters,cycloalkyl esters, lower alkenyl esters, benzyl esters, mono- ordisubstituted lower alkyl esters such as lower omega-(amino, mono- ordialkylamino, carboxy, lower alkoxycarbonyl)—alkyl esters or such aslower alpha-(alkanoyloxy, alkoxycarbonyl or dialkylaminocarbonyl)—alkylesters; conventionally, pivaloyloxymethyl esters and similar esters areused as such.

Because of the close relationship between a free compound, a prodrugderivative and a salt compound, a particular compound in this inventionalso includes its prodrug derivative and salt form, where this ispossible and appropriate.

The compounds of formula (I), preferably of formulae (IA) and (IB), andtheir pharmaceutically acceptable salts have an inhibitory effect on thenatural enzyme renin. The latter passes from the kidneys into the bloodand there brings about the cleavage of angiotensinogen to form thedecapeptide angiotensin I which is then cleaved in the lung, the kidneysand other organs to the octapeptide angiotensin II. Angiotensin IIraises the blood pressure both directly by arterial constriction, andindirectly by releasing the hormone aldosterone, which retains sodiumions, from the adrenals, which is associated with an increase in theextracellular fluid volume. This increase is attributable to the effectof angiotensin II itself or of the heptapeptide angiotensin III formedtherefrom as cleavage product. Inhibitors of the enzymatic activity ofrenin bring about a reduction in the formation of angiotensin I and, asa consequence thereof, the formation of a smaller amount of angiotensinII. The reduced concentration of this active peptide hormone is thedirect cause of the blood pressure-lowering effect of renin inhibitors.

The effect of renin inhibitors is detected inter alia experimentally bymeans of in vitro tests where the reduction in the formation ofangiotensin I is measured in various systems (human plasma, purifiedhuman renin together with synthetic or natural renin substrate). Thefollowing in vitro test of Nussberger et al. (1987) J. CardiovascularPharmacol., Vol. 9, pp. 39-44, is used inter alia. This test measuresthe formation of angiotensin I in human plasma. The amount ofangiotensin I formed is determined in a subsequent radioimmunoassay. Theeffect of inhibitors on the formation of angiotensin I is tested in thissystem by adding various concentrations of these substances. The IC₅₀ isdefined as the concentration of the particular inhibitor which reducesthe formation of angiotensin I by 50%. The compounds of the presentinvention show inhibitory effects in the in vitro systems at minimalconcentrations of about 10⁻⁶ mol/l to about 10⁻¹⁰ mol/l.

Illustrative of the invention, the compounds of examples 8 and 10inhibit the formation of angiotensin I with IC₅₀ values in the range ofabout 0.1-10·10⁻⁹ mol/l.

Renin inhibitors bring about a fall in blood pressure in salt-depletedanimals. Human renin differs from renin of other species. Inhibitors ofhuman renin are tested using primates (marmosets, Callithrix jacchus)because human renin and primate renin are substantially homologous inthe enzymatically active region. The following in vivo test is employedinter alia: the test compounds are tested on normotensive marmosets ofboth sexes with a body weight of about 350 g, which are conscious,unrestrained and in their normal cages. Blood pressure and heart rateare measured with a catheter in the descending aorta and are recordedradiometrically. Endogenous release of renin is stimulated by combininga low-salt diet for 1 week with a single intramuscular injection offurosemide (5-(aminosulfonyl)-4-chloro-2-[(2-furanylmethyl)amino]benzoicacid) (5 mg/kg). 16 h after the furosemide injection, the testsubstances are administered either directly into the femoral artery bymeans of a hypodermic needle or as suspension or solution by gavage intothe stomach, and their effect on blood pressure and heart rate isevaluated. The compounds of the present invention have a bloodpressure-lowering effect in the described in vivo test with i.v. dosesof about 0.003 to about 0.3 mg/kg and with oral doses of about 0.3 toabout 30 mg/kg.

The blood pressure-reducing effect of the compounds described herein canbe tested in vivo using the following protocol:

The investigations take place in 5 to 6-week old, male double transgenicrats (dTGR), which overexpress both human angiotensinogen and humanrenin and consequently develop hypertension (Bohlender J. et al., J. Am.Soc. Nephrol. 2000; 11: 2056-2061). This double transgenic rat strainwas produced by crossbreeding two transgenic strains, one for humanangiotensinogen with the endogenous promoter and one for human reninwith the endogenous promoter. Neither single transgenic strain washypertensive. The double transgenic rats, both males and females,develop severe hypertension (mean systolic pressure, approximately 200mm Hg) and die after a median of 55 days if untreated. The fact thathuman renin can be studied in the rat is a unique feature of this model.Age-matched Sprague-Dawley rats serve as non-hypertensive controlanimals. The animals are divided into treatment groups and receive testsubstance or vehicle (control) for various treatment durations. Theapplied doses for oral administration may range from 0.5 to 100 mg/kgbody weight.

Throughout the study, the animals receive standard feed and tap water adlibitum. The systolic and diastolic blood pressure, and the heart rateare measured telemetrically by means of transducers implanted in theabdominal aorta, allowing the animals free and unrestricted movement.

The effect of the compounds described herein on kidney damage(proteinuria) can be tested in vivo using the following protocol:

The investigations take place in 4-week old, male double transgenic rats(dTGR), as described above. The animals are divided into treatmentgroups and receive test substance or vehicle (control) each day for 7weeks. The applied doses for oral administration may range from 0.5 to100 mg/kg body weight. Throughout the study, the animals receivestandard feed and tap water ad libitum. The animals are placedperiodically in metabolism cages in order to determine the 24-hoururinary excretion of albumin, diuresis, natriuresis, and urineosmolality. At the end of the study, the animals are sacrificed and thekidneys and hearts may also be removed for determining the weight andfor immunohistological investigations (fibrosis, macrophage/T cellinfiltration, etc.).

The bioavailability of the compounds described herein can be tested invivo using the following protocol:

The investigations take place in pre-catheterized (carotid artery) malerats (300 g±20%) that can move freely throughout the study. The compoundis administered intravenously and orally (gavage) in separate sets ofanimals. The applied doses for oral administration may range from 0.5 to50 mg/kg body weight; the doses for intravenous administration may rangefrom 0.5 to 20 mg/kg body weight. Blood samples are collected throughthe catheter before compound administration and over the subsequent24-hour period using an automated sampling device (AccuSampler, DiLabEurope, Lund, Sweden). Plasma levels of the compound are determinedusing a validated LC-MS analytical method. The pharmacokinetic analysisis performed on the plasma concentration-time curves after averaging allplasma concentrations across time points for each route ofadministration. Typical pharmacokinetics parameters to be calculatedinclude: maximum concentration (C_(max)), time to maximum concentration(t_(max)), area under the curve from 0 hours to the time point of thelast quantifiable concentration (AUC_(0-t)), area under the curve fromtime 0 to infinity (AUC_(0-inf)), elimination rate constant (K),terminal half-life (t_(1/2)), absolute oral bioavailability or fractionabsorbed (F), clearance (CL), and Volume of distribution during theterminal phase (Vd).

Five major metabolizing CYP450 enzymes CYP1A2, CYP2C9, CYP2C19, CYP2D6,and CYP3A4 are responsible for more than 95% of the drug metabolizingactivity in humans.

The goals in evaluating in vitro drug metabolism are:

(1) to identify all of the major metabolic pathways that affect the testcompound and its metabolites, including the identification of thespecific enzymes responsible for metabolism and elucidation of theintermediates formed; and(2) to explore and anticipate the effects of the test drug on themetabolism of other drugs and the effects of other drugs on itsmetabolism.

The most complete picture for hepatic metabolism can be obtained withintact liver systems (e.g. hepatocytes, microsomes), in which thecofactors are self-sufficient and the natural orientation and locationfor linked enzymes is preserved.

However, when many compounds have to be tested simultaneously, a simplerscreening tool is advantageous. The cDNAs for the common CYP450s havebeen cloned and the recombinant human enzymatic proteins have beenexpressed in a variety of cells. Use of these recombinant enzymesprovides an excellent way to quickly assess specific enzyme inhibitionactivities and/or confirm results identified in microsomes.

The metabolic properties (inhibition constants on human cytochrome P450isoforms) of the compounds described herein can be tested in vivo usingthe following protocol:

To assess the inhibitory activity towards CYP450 enzymes, the enzymaticreaction is monitored in the presence of different concentrations oftest compound (serial dilution) and compared to maximal enzyme activity(control: no test compound). In principle, inhibition can occur by threedifferent mechanisms: (1) competitive inhibition, (2) non-competitiveinhibition, and (3) mechanism-based inhibition.

In any case, the inhibition strength is dependent on the concentrationof test compound. Testing the CYP450 enzyme activity over a testcompound concentration range identifies the test compound concentrationat which half maximal enzyme inhibition is observed (IC₅₀concentration).

For screening purposes, the inhibitory potential of a test compound canbe tested with ready to use kits (CYP450 High Throughput InhibitorScreening kit, e.g. CYP1A2/CEC, #459500, BD Biosciences, Franklin Lakes,N.J. USA), which are available for all of the five above-mentioned majorCYP isoforms. In such kits, recombinant human CYP450 isoforms expressedin insect cells are incubated with isoform specific, fluorogenicsubstrates in the presence of different test compound concentrations.Enzymatic activity converts the fluorogenic substrate into afluorochrome product, the concentration of which is measured with afluorospectrophotometer. Fluorescence is directly proportional to enzymeactivity.

In a typical standard assay using the CYP450 High Throughput InhibitorScreening kit, a compound is tested at 2 nM to 33 μM concentration rangein a phosphate buffer (50 mM, pH 7.4) containing a glucose 6-phosphatedehydrogenase/NADP/NADPH regeneration system and a suitable fluorogenicsubstrate: e.g. 3-cyano-7-ethoxycoumarin (CYP1A2). As controlinhibitors, the following substances can be used: furafylline (CYP1A2),sulfaphenazole (CYP2C9), tranylcypromine (CYP2C19), quinidine (CYP2D6)and ketoconazole (CYP3A4).

The reaction is started by the addition of 2.5 nM (final concentration)CYP450 isozyme, incubated at 37° C. for 15 to 45 minutes, and thenterminated by the addition of 187.5 mM tris-hydroxy-aminomethanebase/acetonitrile (20/80, v/v).

The amount of generated fluorochrome is then determined by fluorescencespectroscopy with suitable excitation and emission wavelength settings:e.g. 410 nm excitation and 460 nm emission wavelength (CYP1A2).

Alternatively and/or complimentary, assays using human liver microsomes(e.g. BD Biosciences, #452161) in combination with a CYPisoform-specific standard substrate (e.g. midazolam for CYP3A4/5) asdescribed by R. L. Walsky and R. S. Obach in Validated assay for humancytochrome p450 activities; Pharmacokinetics, Pharmacodynamics, and DrugMetabolism, Pfizer, Groton, Conn.; Drug Metabolism and Disposition:(2004)32, 647-660, can be used. To determine whether a test compoundinhibits CYP3A enzyme activity, for example, hydroxylation of midazolamby human liver microsomes at varying test compound concentrations ismonitored. Hydroxy-midazolam production is directly proportional toenzyme activity and can be determined by liquid chromatography-tandemmass spectrometry. Additionally, the microsomal assay can be run withoutand with a 15 min pre-incubation of microsomes with test compound priorto the addition of standard substrate. Test compounds or theirmetabolite(s) that have the potential to irreversibly modify the P450enzyme will have a stronger inhibitory effect after pre-incubation.

In a typical standard assay using the human liver microsome assay,compounds are tested at 10 nM to 50 μM concentration range in aphosphate buffer (100 mM potassium phosphate, 3.3 mM MgCl₂, pH 7.4)containing a NADPH regeneration system (glucose 6-phosphatedehydrogenase, NADP, NADPH) and 10 μM substrate (e.g. midazolam forCYP3A4/5) and 0.1 mg/mL microsomal protein. As control inhibitors, thesame substances as described above can be used (e.g. ketoconazole(CYP3A4/5)). If pre-incubation of the compound is desired, all assaycomponents except substrate are mixed and incubated for 15 minutes at37° C. After that period, substrate is added to the assay mix and thenincubation at 37° C. is continued for 15 minutes. Withoutpre-incubation, all assay components are mixed simultaneously and thenincubated at 37° C. for 15 minutes. Termination of the enzymaticreaction is achieved by the addition of a HCOOH/acetonitrile/H₂O (Apr.30, 1966, v/v/v) solution. Samples are then incubated in therefrigerator (4±2° C.) for 1 h±10 min to increase protein precipitation.Directly before analysis by LC/MSMS, the samples are centrifuged at3,500 g for 60 min at 4° C. to separate precipitated protein. Thesupernatant is mixed with acetonitrile/water (50/50, v/v), and thendirectly analyzed for compound content with LC/MSMS.

Evaluation of the data from either experimental setup is then done asfollows: the fraction of remaining activity at a specific compoundconcentration versus the activity in the control as a function ofcompound concentration is used to compute IC₅₀ values. This is done byfitting a 4-parameter logistic function to the experimental data set.

The compounds of the formula (I), preferably of formulae (IA) and (IB),and their pharmaceutically acceptable salts can be used as medicines,e.g. in the form of pharmaceutical compositions. The pharmaceuticalcompositions can be administered enterally, such as orally, e.g. in theform of tablets, lacquered tablets, sugar-coated tablets, hard and softgelatine capsules, solutions, emulsions or suspensions, nasally, e.g. inthe form of nasal sprays, rectally, e.g. in the form of suppositories,or transdermally, e.g. in the form of ointments or patches,ophtalmologically, e.g. in the form of solutions, suspensions,ointments, gels, pulmonary, e.g. in the form of pulmonary aerosols or toother mucosal tissues. However, administration is also possibleparenterally, such as intramuscularly or intravenously, e.g. in the formof solutions for injection.

Tablets, lacquered tablets, sugar-coated tablets and hard gelatinecapsules can be produced by processing the compounds of the formula (I)and their pharmaceutically acceptable salts with pharmaceutically inertinorganic or organic excipients. Excipients of these types which can beused for example for tablets, sugar-coated tablets and hard gelatinecapsules are lactose, maize starch or derivatives thereof, talc, stearicacid or salts thereof etc.

Excipients suitable for soft gelatine capsules are, for example,vegetable oils, waxes, fats, semisolid and liquid polyols etc.

Excipients suitable for producing solutions and syrups are, for example,water, polyols, sucrose, invert sugar, glucose etc.

Excipients suitable for solutions for injection are, for example, water,alcohols, polyols, glycerol, vegetable oils, bile acids, lecithin etc.

Excipients suitable for suppositories are, for example, natural orhardened oils, waxes, fats, semiliquid or liquid polyols etc.

The pharmaceutical compositions may in addition comprise preservatives,solubilizers, viscosity-increasing substances, stabilizers, wettingagents, emulsifiers, sweeteners, colorants, aromatizers, salts to alterthe osmotic pressure, buffers, coating agents or antioxidants. They mayalso comprise other substances of therapeutic value.

The present invention further provides the use of the compounds of theformula (I), preferably of formulae (IA) and (IB), and theirpharmaceutically acceptable salts in the treatment, for the delay ofprogression or for the prevention of high blood pressure, heart failure,glaucoma, myocardial infarction, renal failure, diabetic nephropathy orrestenoses.

The present invention also provides a method in the treatment, for thedelay of progression or for the prevention of high blood pressure, heartfailure, glaucoma, myocardial infarction, renal failure, diabeticnephropathy or restenoses comprising administering a therapeuticallyeffective amount of the compounds of the formula (I), preferably offormulae (IA) and (IB), and their pharmaceutically acceptable salts to asubject in need thereof.

The subject/patient may be an animal, mammal, a placental mammal, amarsupial (e.g., kangaroo, wombat), a monotreme (e.g., duckbilledplatypus), a rodent (e.g., a guinea pig, a hamster, a rat, a mouse),murine (e.g., a mouse), a lagomorph (e.g., a rabbit), avian (e.g., abird), canine (e.g., a dog), feline (e.g., a cat), equine (e.g., ahorse), porcine (e.g., a pig), ovine (e.g., a sheep), bovine (e.g., acow), a primate, simian (e.g., a monkey or ape), a monkey (e.g.,marmoset, baboon), an ape (e.g., gorilla, chimpanzee, orangutan,gibbon), or a human.

The compounds of the formula (I), preferably of formulae (IA) and (IB),and their pharmaceutically acceptable salts can also be administered incombination with one or more agents having cardiovascular activity, e.g.alpha- and beta-blockers such as phentolamine, phenoxybenzamine,prazosin, terazosin, tolazine, atenolol, metoprolol, nadolol,propranolol, timolol, carteolol etc.; vasodilators such as hydralazine,minoxidil, diazoxide, nitroprusside, flosequinan etc.; calciumantagonists such as amrinone, bencyclan, diltiazem, fendiline,flunarizine, nicardipine, nimodipine, perhexyline, verapamil,gallopamil, nifedipine etc.; ACE inhibitors such as cilazapril,captopril, enalapril, lisinopril etc.; potassium activators such aspinacidil; antiserotoninergics such as ketanserine; thromboxanesynthetase inhibitors; neutral endopeptidase inhibitors (NEPinhibitors); angiotensin II antagonists; and diuretics such ashydrochlorothiazide, chlorothiazide, acetazolamide, amiloride,bumetanide, benzthiazide, ethacrynic acid, furosemide, indacrinone,metolazone, spironolactone, triamterene, chlorthalidone etc.;sympatholytics such as methyldopa, clonidine, guanabenz, reserpine; andother agents suitable for the treatment of high blood pressure, heartfailure or vascular disorders associated with diabetes or renaldisorders such as acute or chronic renal failure in humans and animals.Such combinations can be used separately or in products which comprise aplurality of components.

Further substances which can be used in combination with the compoundsof formula (I), preferably of formulae (IA) and (IB), are the compoundsof classes (i) to (ix) on page 1 of WO 02/40007 (and the preferences andexamples detailed further therein) and the substances mentioned on pages20 and 21 of WO 03/027091.

The dosage may vary within wide limits and must of course be adapted tothe individual circumstances in each individual case. In general, adaily dose appropriate for oral administration ought to be from about 3mg to about 3 g, preferably about 10 mg to about 1 g, e.g. approximately300 mg per adult person (70 kg), divided into preferably 1-3 singledoses, which may be for example of equal size, although the stated upperlimit may also be exceeded if this proves to be indicated, and childrenusually receive a reduced dose appropriate for their age and bodyweight.

The compounds of the formula (I), preferably of formulae (IA) and (IB),and their pharmaceutically acceptable salts can also be administeredwith one or several varying dosing intervals, as long as the intendedtherapeutic effect is sustained or as long as further therapeuticintervention is not required.

EXAMPLES

The examples which follow illustrate the present invention. Alltemperatures are reported in degrees Celsius, pressures in mbar. Unlessstated otherwise, the reactions take place at RT. The abbreviation“Rf=xx (A)” means, for example, that the Rf value xx is obtained in thesolvent system A. The ratio of the solvents relative to one another isalways reported in parts by volume. Chemical names of end products andintermediates were obtained with the aid of the program AutoNom 2000(Automatic Nomenclature).

HPLC gradient on Hypersil BDS C-18 (5 μm); column: 4×125 mm

-   I 90% H₂O*/10% CH₃CN* to 0% H₂O*/100% CH₃CN* in 5 min+2.5 min (1.5    ml/min)-   II 95% H₂O*/5% CH₃CN* to 0% H₂O*/100% CH₃CN* in 30 min+5 min (0.8    ml/min)    * contains 0.1% trifluoroacetic acid

The following abbreviations are used:

-   Rf ratio of distance which a substance travels to distance of the    eluent front from the start point in thin layer chromatography-   Rt retention time of a substance in HPLC (in minutes)-   m.p. melting point (temperature)

The following abbreviations are used:

-   AcOH acetic acid-   n-BuLi n-butyllithium-   t-BuOH tert-butanol-   CH₂Cl₂ dichloromethane-   CHCl₃ chloroform-   CH₃CN acetonitrile-   Cs₂CO₃ caesium carbonate-   Cy cyclohexane-   DCC dicyclohexylcarbodiimide-   DIBAL diisobutylaluminium hydride-   DMA dimethylacetamide-   4-DMAP 4-dimethylamino pyridine-   DME 1,2-dimethoxyethane-   DMF N,N-dimethylformamide-   DMPU 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone-   dppf 1,1′-bis(diphenylphosphino)-ferrocene [12150-46-8]-   EDC.HCl N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride    [25952-53-8]-   Et₃N triethylamine-   Et₂O diethylether-   EtOAc ethyl acetate-   EtOH ethanol-   h hour-   HCl hydrochloric acid-   H₂O water-   K₂CO₃ potassium carbonate-   LiBH₄ lithium borohydride-   LiCl lithium chloride-   MeI methyl iodide-   MeOH methanol-   min minute(s)-   m.p. melting point (temperature)-   N₂ nitrogen-   Na₂CO₃ sodium carbonate-   NaH sodium hydride-   NaHCO₃ sodium bicarbonate-   Na₂HPO₄ di-sodium hydrogen phosphate-   NaOH sodium hydroxide-   Na₂SO₄ sodium sulfate-   NH₃ ammonia-   NH₄Br ammonium bromide-   NH₄Cl ammonium chloride-   NH₄OH ammonium hydroxide-   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium [51364-51-3]-   Pd(PPh₃)₄ tetrakis-triphenylphosphine palladium(0)-   P(tert-Bu)₃ tri-tert-butylphosphine-   Ra/Ni Raney-nickel-   Rf ratio of distance which a substance travels to distance of the    eluent front from the start point in thin layer chromatography-   Rt retention time of a substance in HPLC (in minutes)-   RT room temperature corresponding to about 23° C.-   TBACl tetrabutyl ammonum chloride-   TBAF tetrabutyl ammonum fluoride-   TBAI tetrabutyl ammonum iodide-   TBME tert-butyl methyl ether-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TMS trimethylsilyl

Example 16-{(3R,4R,6S)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-[(R)-2-(1H-tetrazol-5-yl)-propyl]-piperidin-3-yloxymethyl}-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

The title compound is prepared according to the general procedure Astarting from6-[(3R,4R,6S)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-6-[(R)-2-(1H-tetrazol-5-yl)-propyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineand is identified based on its Rf-value.

The starting material(s) is (are) prepared as follows:

a)6[(3R,4R,6S)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-[(R)-2-(1H-tetrazol-5-yl)-propyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a stirred solution of 1.0 mmol of(R)-3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2-methyl-propionitrileand 0.3 mmol of dibutyltin oxide in 10 ml of toluene is added 40.0 mmolof trimethylsilylazide. The reaction mixture is heated to 125° C.overnight, concentrated under reduced pressure and the residue ispurified by flash chromatography (SiO₂ 60 F) to afford the titlecompound which is identified based on its Rf-value.

b)(R)-3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2-methyl-propionitrile

The title compound is prepared according to the general procedure Gstarting from methanesulfonic acid(S)-2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethylester and is identified based on its Rf-value.

c) Methanesulfonic acid(S)-2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethylester

The title compound is prepared according to the general procedure Fstarting from(S)-1-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propan-2-oland is identified based on its Rf-value.

d)(S)-1-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propan-2-ol

To a stirred solution of 1.0 mmol of1-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propan-2-onein 10 ml of THF are added 2.0 mmol of borane tetrahydrofuran complex (1Min THF) at RT. The reaction mixture is stirred for 3 h, quenched with 60ml of MeOH and concentrated under reduced pressure. The resultingdiastereomeric mixture is separated by flash chromatography (SiO₂ 60 F)to afford the title compound which is identified based on its Rf-value.

e)1-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propan-2-one

To a stirred solution of 1.0 mmol ofN-methoxy-2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-N-methyl-acetamidein 10 ml of THF are added 1.2 mmol of methyl magnesium bromide (3M inTHF) at 0° C. The reaction mixture is stirred for 1 h, diluted with anaqueous solution of 1N potassium hydrogen sulfate, extracted with TBME.The organic phases are combined and dried over Na₂SO₄. The residue ispurified by flash chromatography (SiO₂ 60 F) to afford the titlecompound which is identified based on its Rf-value.

f)N-Methoxy-2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-N-methyl-acetamide

According to general procedure D, 1.0 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid are reacted with N,O-dimethylhydroxylamine hydrochloride to affordthe title compound which is identified based on its Rf-value.

g)[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid

The title compound is prepared according to general procedure H startingfrom 1.0 mmol of[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetonitrile.Rf=0.15 (EtOAc); Rt=5.17 (gradient I).

h)[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetonitrile

The title compound is prepared according to general procedure G startingfrom 1.0 mmol of methanesulfonic acid(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-ylmethylester. Rf=0.48 (EtOAc/heptane 2:1); Rt=5.57 (gradient I).

i) Methanesulfonic acid(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-ylmethylester

The title compound is prepared according to general procedure F startingfrom 1.0 mmol of[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-methanol.Rf=0.45 (EtOAc/heptane 2:1); Rt=5.52 (gradient I).

j)[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-methanol

To a stirred solution of 1.0 mmol of6-[(3R,4R,6S)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-6-triisopropylsilanyloxymethyl-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazinein 10 ml of THF are added 2.5 mmol of a 1N solution of TBAF in THF atRT. The reaction mixture is stirred for 2 h at RT, diluted with waterand extracted with TBME. The combined organic phases are dried overNa₂SO₄ and concentrated under reduced pressure. The crude product ispurified by flash chromatography (SiO₂ 60 F) to afford the titlecompound as a yellow oil. Rf=0.29 (EtOAc/heptane 3:1); Rt=5.26 (gradientI).

k)6-[(3R,4R,6S)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-6-triisopropylsilanyloxymethyl-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a solution of 1.0 mmol of6-[(3R,4R,6S)-4-(4-chloromethyl-phenyl)-1-(toluene-4-sulfonyl)-6-triisopropylsilanyloxymethyl-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine,1.5 mmol of (R)-3-methoxy-2-methyl-propan-1-ol [911855-78-2] and 0.1mmol of TBAI in 5 ml of DMF are added 1.65 mmol of NaH (55% indispersion oil) at 0° C. The reaction mixture is allowed to warm to RTand stirred overnight, diluted with water, extracted with TBME and theorganic phase is dried over Na₂SO₄. The organic phase is concentratedunder reduced pressure and the residue is purified by flashchromatography (SiO₂ 60 F) to afford the title compound as a pale yellowoil. Rf=0.20 (EtOAc/heptane 1:2).

l)6-[(3R,4R,6S)-4-(4-Chloromethyl-phenyl)-1-(toluene-4-sulfonyl)-6-triisopropylsilanyloxymethyl-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a solution of 1.0 mmol of{4-[(2S,4R,5R)-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-2-triisopropylsilanyloxymethyl-piperidin-4-yl]-phenyl}-methanol[911707-31-8], 1.2 mmol of Et₃N and 0.1 mmol of TBAClin 10 ml of CH₂Cl₂are added 1.1 mmol of methanesulfonyl chloride at 0° C. The reactionmixture is allowed to warm to RT and stirred overnight, diluted withCH₂Cl₂, washed with saturated aqueous NaHCO₃, water and brine. Theorganic phase is dried over Na₂SO₄ and is concentrated under reducedpressure to afford the title compound as a yellow oil. Rf=0.77(EtOAc/heptane 2:1).

Example 2N,N-Diethyl-2-{(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-piperidin-2-yl}-acetamide

The title compound is prepared according to the general procedure Astarting fromN,N-diethyl-2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetamideand is identified based on its Rf-value.

The starting material(s) is (are) prepared as follows:

a)N,N-Diethyl-2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetamide

According to general procedure D, 1.0 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid (example 1g) are reacted with diethylamine to afford the titlecompound which is identified based on its Rf-value.

Example 3N-((S)-2-{(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-piperidin-2-yl}-1-methyl-ethyl)-acetamide

The title compound is prepared according to the general procedure Astarting fromN-{(S)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethyl}-acetamideto afford the title compound as a colourless oil. Rf=0.45(CH₂Cl₂/MeOH/25% conc. NH₃ 200:20:1); Rt=3.90 (gradient I).

The starting material(s) is (are) prepared as follows:

a)N-{(S)-2-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethyl}-acetamideandN-{(R)-2-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethyl}-acetamide

To a solution of 1.0 mmol of (R andS)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethylamine,4.0 mmol of Et₃N in 25 ml of CH₂Cl₂ are added 2.0 mmol of acetylchloride at 0° C. and the resulting mixture is stirred for 1 h. Thereaction mixture is diluted with water and extracted with CH₂Cl₂. Theorganic phases are combined, dried over Na₂SO₄ and concentrated underreduced pressure. The resulting diastereomeric mixture is separated byflash chromatography (SiO₂ 60 F) to afford:

N-{(S)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethyl}-acetamide.Colourless oil; Rf=0.21 (CH₂Cl₂/MeOH/NH₃ 100:2.5:0.05); Rt=21.72(gradient II).N-{(R)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethyl}-acetamide.Colourless oil, Rf=0.22 (CH₂Cl₂/MeOH/NH₃ 100:2.5:0.05); Rt=21.88(gradient II).

b) (R andS)-2-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethylamine

According to general procedure E, 1.0 mmol of (R andS)-6-[(3R,4R,6S)-6-(2-azido-propyl)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineare hydrogenated in 20 ml of MeOH to afford the title compound as adiastereomeric mixture after flash chromatography (SiO₂ 60 F), as ayellow oil. Rf=0.14 (CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 200:10:1); Rt=4.82(gradient I).

c) (R andS)-6-[(3R,4R,6S)-6-(2-Azido-propyl)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a solution of 1.0 mmol of methanesulfonic acid (R andS)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethylester in 5.0 ml DMPU are added 10.0 mmol of sodium azide and the mixtureis heated to 80° C. overnight. The reaction mixture is diluted withwater and extracted with TBME. The organic phases are combined andconcentrated under reduced pressure. The residue is purified by flashchromatography (SiO₂ 60 F) to afford the title compound as colourlessoil. Rf=0.38 (EtOAc/heptane 1:1); Rt=6.00 (gradient I).

d) Methanesulfonic acid (R andS)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethylester

The title compound is prepared according to general procedure F startingfrom 1.0 mmol of (R andS)-1-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)piperidin-2-yl]-propan-2-ol.Rf=0.23 (EtOAc/heptane 2:1); Rt=5.06 (gradient I).

e) (R andS)-1-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propan-2-ol

To a stirred solution of 1.0 mmol of[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetaldehydein 10 ml of THF are added 2.0 mmol of methyl magnesium bromide (3M inTHF) at 0° C. The reaction mixture is allowed to warm to RT and stirredfor 1 h, diluted with an aqueous solution of 1N potassium hydrogensulfate and extracted with TBME. The organic phases are combined anddried over Na₂SO₄. The residue is purified by flash chromatography (SiO₂60 F) to afford the title compound as colourless oil. Rf=0.38(EtOAc/heptane 2:1); Rt=5.35 (gradient I).

f)[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetaldehyde

To 1.0 mmol of[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetonitrile(example 1h) in 10 ml of toluene are added 1.2 mmol of DIBAL (1.5M intoluene) at −40° C. The reaction mixture is stirred for 4 h at −40° C.to −20° C. and poured into 1N aqueous HCl. The organic phase isseparated, washed with brine, dried over Na₂SO₄ and concentrated underreduced pressure. The residue is purified by flash chromatography (SiO₂60 F) to afford the title compound as a colourless oil. Rf=0.69(EtOAc/heptane 3:1); Rt=5.44 (gradient I).

Example 4N-((R)-2-{(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-piperidin-2-yl}-1-methyl-ethyl)-acetamide

The title compound is prepared according to the general procedure Astarting fromN-{(R)-2-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1-methyl-ethyl}-acetamide(example 3a) to afford the title compound as a colourless oil. Rf=0.48(CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 200:20:1); Rt=4.12 (gradient I).

Example 53-{(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-piperidin-2-yl}-2,2,N-trimethyl-propionamide

The title compound is prepared according to the general procedure Astarting from3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2,N-trimethyl-propionamideto afford the title compound as a yellow oil. Rf=0.33(CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 90:10:1); Rt=4.16 (gradient I).

The starting material(s) is (are) prepared as follows:

a)3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2,N-trimethyl-propionamide

According to general procedure D, 1.0 mmol of3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid are reacted with methylamine (8M in EtOH) to afford the titlecompound as yellow oil. Rf=0.25 (EtOAc/heptane 3:1); Rt=5.36 (gradientI).

b)3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid

To a solution of 1.0 mmol of3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid methyl ester in 10 ml of THF and 10 ml of MeOH are added 10 ml of a3M lithium hydroxide solution. The reaction mixture is stirred for 4 hat 65° C. The organic solvents are evaporated under reduced pressure andthe remaining solution is acidified with 3M aqueous HCl until pH 2. Thismixture is extracted with 100 ml of EtOAc (3×). The combined organiclayers are washed with brine and dried with Na₂SO₄. The organic layer isfiltered and evaporated under reduced pressure. The residue is purifiedby flash chromatography (SiO₂ 60 F) to afford the title compound as apale orange foam. Rf=0.28 (EtOAc/heptane/HCOOH 2:1:0.1); Rt=5.50(gradient I).

c)3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid methyl ester

To a solution of 4.0 mmol of methyl isobutyrate in 2 ml of THF at −78°C. are added 4.0 mmol of LDA (0.5 M in THF) and the reaction mixture isstirred for 30 min at −78° C. Then 8.0 mmol of hexamethylphosphoramideare added at −78° C. and the mixture is stirred for 30 min. A solutionof 1.0 mmol of6-[(3R,4R,6S)-6-bromomethyl-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazinein 2 ml of THF is then added dropwise to the “enolate” at −78° C. andthe solution is stirred for 30 min. The reaction mixture is allowed towarm to −12° C. and stirred at this temperature for 40 min. The solutionis quenched with 1M aqueous HCl and extracted with 200 ml of EtOAc (3×).The combined organic layers are washed with brine and dried with Na₂SO₄.The organic layer is filtered and evaporated under reduced pressure. Theresidue is purified by flash chromatography (SiO₂ 60 F) to afford thetitle compound as yellow oil. Rf=0.60 (EtOAc/heptane 3:1); Rt=6.03(gradient I).

d)6-[(3R,4R,6S)-6-Bromomethyl-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a solution of 1.0 mmol of methanesulfonic acid(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-ylmethylester (example 11) in 4 ml of DMF are added 10.0 mmol of lithium bromideand the reaction mixture is stirred for 14 h at 65° C. The reactionmixture is allowed to warm to RT and 100 ml of water are added. Thismixture is extracted with 100 ml of TBME (3×). The combined organiclayers are washed with brine and dried with Na₂SO₄. The organic layer isfiltered and evaporated under reduced pressure. The residue is purifiedby flash chromatography (SiO₂ 60 F) to afford the title compound as acolourless foam. Rf=0.47 (EtOAc/heptane 1:1); Rt=6.07 (gradient I).

Example 63-{(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-piperidin-2-yl}-2,2,N-trimethyl-propionamide

The title compound is prepared according to the general procedure Astarting from3-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2,N-trimethyl-propionamideand is identified based on its Rf-value.

The starting material(s) is (are) prepared as follows:

a)3-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2,N-trimethyl-propionamide

According to general procedure D, 1.0 mmol of3-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid are reacted with methylamine (8M in EtOH) to afford the titlecompound which is identified based on its Rf-value.

b)3-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid

According to the procedure described in example 5b, the title compoundis prepared from3-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid methyl ester and is identified based on its Rf-value.

c)3-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid methyl ester

To a suspension of 4.0 mmol of potassium tert-butoxide and 0.2 mmol of18-crown-6 [17455-13-9] in 25 ml of THF at −78° C. are added 1.0 mmol of3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propionicacid methyl ester in 5 ml of THF and the reaction mixture is stirred for1 h at −78° C. Then 4.1 mmol of MeI in 3 ml of THF are added at −78° C.and the mixture is stirred for 65 minutes at −78° C. The reactionmixture is quenched with 0.5 M aqueous HCl to pH 2 and extracted with200 ml of EtOAc (3×). The combined organic layers are washed with brineand dried with Na₂SO₄. The organic layer is filtered and evaporatedunder reduced pressure. The residue is purified by flash chromatography(SiO₂ 60 F) to afford the title compound which is identified based onits Rf-value.

d)3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propionicacid methyl ester

To a solution of 1.0 mmol of3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propionicacid in 5 ml of MeOH at 0° C. are added 5.0 mmol of TMS-diazomethane (2Min hexane) until the conversion to the methyl ester is complete. Thereaction mixture is treated with magnesium sulfate to destroy the excessof TMS-diazomethane. The solids are removed by filtration and theorganic layer is concentrated under reduced pressure. The residue ispurified by flash chromatography (SiO₂ 60 F) to afford the titlecompound which is identified based on its Rf-value.

e)3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propionicacid

According to the procedure described in example 1g the title compound isprepared from3-[(2S,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propionitrileand is identified based on its Rf-value.

f)3-[(2S,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-propionitrile

According to the procedure described in example 1h the title compound isprepared from methanesulfonic acid2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethylester and is identified based on its Rf-value.

g) Methanesulfonic acid2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethylester

According to the procedure described in example 11 the title compound isprepared from2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethanoland is identified based on its Rf-value.

h)2-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethanol

To a stirred solution of 1.0 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid (example 1g) in 4 ml of THF are added 2.2 mmol of boranetetrahydrofuran complex (1M in THF) at RT. The reaction mixture isstirred for 3 h, quenched with 60 ml of MeOH and concentrated underreduced pressure. The residue is purified by flash chromatography (SiO₂60 F) to afford the title compound which is identified based on itsRf-value.

Example 7N-(2-{(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-piperidin-2-yl}-1,1-dimethyl-ethyl)-acetamide

The title compound is prepared according to the general procedure Astarting fromN-{2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1,1-dimethyl-ethyl}-acetamideand is identified based on its Rf-value.

The starting material(s) is (are) prepared as follows:

a)N-{2-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1,1-dimethyl-ethyl}-acetamide

To a solution of 1.0 mmol of2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1,1-dimethyl-ethylamine,3.0 mmol of Et₃N in 10 ml of CH₂Cl₂ are added 1.5 mmol of acetylchloride at 0° C. and the resulting mixture is stirred for 1 h. Thereaction mixture is diluted with water and extracted with CH₂Cl₂. Theorganic phases are combined, dried over Na₂SO₄ and concentrated underreduced pressure. The residue is purified by flash chromatography (SiO₂60 F) to afford the title compound which is identified based on itsRf-value.

b)2-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1,1-dimethyl-ethylamine

The title compound is prepared according to the general procedure Estarting from{2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1,1-dimethyl-ethyl}-carbamicacid benzyl ester and is identified based on its Rf-value.

c){2-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-1,1-dimethyl-ethyl}-carbamicacid benzyl ester

A mixture of 1.0 mmol of6-[(3R,4R,6R)-6-(2-isocyanato-2-methyl-propyl)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineand 20.0 mmol of benzyl alcohol is heated at 115° C. for 14 h. Thereaction mixture is directly purified by flash chromatography (SiO₂ 60F) to afford the title compound which is identified based on itsRf-value.

d)6-[(3R,4R,6R)-6-(2-Isocyanato-2-methyl-propyl)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 1.0 mmol of3-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionylazide in 10 ml of toluene is heated at 115° C. for 1.5 h. The reactionmixture is concentrated under reduced pressure to afford the crude titlecompound which is identified based on its Rf-value.

e)3-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionylazide

To a solution of 1.0 mmol of3-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-2,2-dimethyl-propionicacid (example 6b) and 2.5 mmol of Et₃N in 10 ml of THF are added 2.0mmol of ethyl chloroformate at 0° C. and the mixture is stirred for 1 hat 0° C. Then a solution of 20.0 mmol of sodium azide in 5 ml of wateris added dropwise and the reaction mixture is stirred at 0° C. for 45min. The mixture is diluted with 2 ml of water. This mixture isextracted with 50 ml of EtOAc (3×), the combined organic layers arewashed with water (2×) and dried with Na₂SO₄. The organic layer isfiltered and evaporated under reduced pressure to afford thecorresponding crude acyl azide which is identified based on itsRf-value.

Example 86-[(3R,4R,6R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-(4-methyl-4H-[1,2,4]triazol-3-ylmethyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

According to general procedure A, 1 mmol of6-[(3R,4R,6R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-6-(4-methyl-4H-[1,2,4]triazol-3-ylmethyl)-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineis used to afford the title compound as a yellow oil. Rf=0.15(CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 200:20:1); Rt=15.20 (gradient II).

The starting materials are prepared as follows:

a)6-[(3R,4R,6R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-(4-methyl-4H-[1,2,4]triazol-3-ylmethyl)-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

Into a solution of 1 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid [1-dimethylamino-meth-(E)-ylidene]-hydrazide in 2 ml ofacetonitrile is condensed ca. 1 ml of gaseous methylamine, at RT. AcOH(2 ml) is carefully added, then the reaction system is sealed, andheated at 100° C. for 4 h. The reaction mixture is cooled to RT, pouredinto saturated aqueous NaHCO₃, and extracted with EtOAc (3×). Thecombined organic extracts are dried over Na₂SO₄, concentrated underreduced pressure, and purified by flash chromatography (SiO₂ 60 F), toafford the title compound as a viscous brown-orange oil. Rf=0.43(CH₂Cl₂/MeOH/25% conc. NH₃ 200:20:1); Rt−=4.84 (gradient I).

b)[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid [1-dimethylamino-meth-(E)-ylidene]-hydrazide

A solution of 1 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid hydrazide in 5 ml of CH₂Cl₂ is treated at RT with 1.1 mmol ofN,N-dimethylformamide dimethylacetal and refluxed for 1 h. The mixtureis cooled to RT, and concentrated under reduced pressure to afford thetitle compound as a viscous brown-orange oil. Rt=4.75 (gradient I).

c)[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid hydrazide

A solution of 1 mmol ofN′-{2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetyl}-hydrazinecarboxylicacid tert-butyl ester in 13 ml of CH₂Cl₂ is treated at 0° C. with 3.3 mlof trifluoroacetic acid. The reaction mixture is stirred at 0° C. for 1h, then at RT for 3 h and diluted with CH₂Cl₂, basified with saturatedaqueous NaHCO₃. The aqueous layer is extracted with CH₂Cl₂ (2×), thecombined organic extracts are washed with H₂O, then with brine, driedover Na₂SO₄, and concentrated under reduced pressure, affording thetitle compound as an orange oil. Rf=0.45 (CH₂Cl₂/MeOH/25%)/0 conc. NH₃200:20:1); Rt=4.57 (gradient I).

d)N′-{2-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-acetyl}-hydrazinecarboxylicacid tert-butyl ester

A solution of 1 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid (example 1g) in 5 ml of CH₂Cl₂ is treated at RT with 1.5 mmol of1-chloro-N,N,2-trimethylpropenylamine, and stirred at RT for 2.5 h. Thissolution is added slowly, at 0° C., to a solution of 5 mmol oftert-butyl carbazate [870-46-2] and 5 mmol of Et₃N in 1 ml of CH₂Cl₂.The reaction mixture is stirred at 4° C. for 16.5 h, then quenched withH₂O. The aqueous layer is extracted with CH₂Cl₂ (2×). The combinedorganic extracts are washed with brine, dried over Na₂SO₄, andconcentrated under reduced pressure. The residue is purified by flashchromatography (SiO₂ 60 F) to afford the title compound as a yellow oil.Rf=0.34 (EtOAc/Heptane 2:1); Rt=5.28 (gradient I).

Example 96-[(3R,4R,6R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-(2-[1,2,4]triazol-4-yl-ethyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

According to general procedure A, 1 mmol of6-[(3R,4R,6R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-6-(2-[1,2,4]triazol-4-yl-ethyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineis used to afford the title compound as a yellow oil. Rf=0.25(CH₂Cl₂/MeOH/25% conc. NH₃ 90:10:1); Rt=3.95 (gradient I).

The starting material(s) is (are) prepared as follows:

a)6-[(3R,4R,6R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-6-(2-[1,2,4]triazol-4-yl-ethyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a solution of 1 mmol of2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethylamineand 3.5 mmol of N,N-dimethylformamidazine [16114-05-9] in 5 ml oftoluene is treated at RT with 0.1 mmol of p-toluenesulfonic acid. Thereaction mixture is stirred at 80° C. for 20 h, cooled to RT, anddiluted with EtOAc. The organic phase is washed sequentially with 1Ncitric acid, saturated aqueous NaHCO₃ and brine, dried over Na₂SO₄, andconcentrated under reduced pressure. The residue is purified by flashchromatography (SiO₂, 60 F), to afford the title compound as acolourless resin. Rf=0.22 (EtOAc/MeOH 9:1); Rt=4.83 (gradient I).

b)2-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethylamine

According to general procedure E, 1 mmol of6-[(3R,4R,6R)-6-(2-azido-ethyl)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineis reacted in MeOH, to afford the title compound as a yellow oil.Rf=0.20 (CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 200:10:1); Rt=4.67 (gradient I).

c)6-[(3R,4R,6R)-6-(2-Azido-ethyl)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 1 mmol of methanesulfonic acid2-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-ethylester (example 6g) and 10 mmol of sodium azide in 1.5 ml of DMPU isstirred at 80° C. for 3 h. The solution is cooled to RT, diluted with 3ml of TBME, filtered through SiO₂ 60 F, eluting with EtOAc-Heptane 1:1.The filtrate is concentrated under reduced pressure and purified byflash chromatography (SiO₂ 60 F), to afford the title compound as acolourless resin. Rf=0.54 (EtOAc/heptane 2:1); Rt=5.90 (gradient I).

Example 106-{(3R,4R,6R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-morpholin-4-ylmethyl-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

According to general procedure A, 1 mmol of6-[(3R,4R,6R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-6-morpholin-4-ylmethyl-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineis used to afford the title compound as a yellow oil. Rf=0.37(CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 200:20:1); Rt=3.77 (gradient I).

The starting materials are prepared as follows:

a)6-[(3R,4R,6R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-6-morpholin-4-ylmethyl-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

A solution of 1 mmol ofC-[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-methylaminein 7 ml of DMSO at RT is treated with 4 mmol of Et₃N, followed by 1.2mmol of (bis-2-bromoethyl)ether [4497-29-4]. The reaction mixture isstirred at 40° C. for 16 h, cooled to RT, and diluted with EtOAc. Theorganic phase is washed with H₂O (2×). The combined aqueous phases areextracted with EtOAc. The combined organic extracts are dried overNa₂SO₄ and concentrated under reduced pressure. The residue is purifiedby flash chromatography (SiO₂, 60 F), to afford the title compound as anorange oil. Rf=0.39 (EtOAc); Rt=4.91 (gradient I).

b)C-[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-methylamine

According to general procedure E, 1 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-ylmethyl]-carbamicacid benzyl ester is reacted in MeOH, to afford the title compound as ayellow oil. Rf=0.1 (CH₂Cl₂/MeOH/25%)/0 conc. NH₃ 200:10:1); Rt=4.62(gradient I).

c)[(2R,4R,5R)-4-[4-((S)-3-Methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-ylmethyl]-carbamicacid benzyl ester

A solution of 1 mmol of6-[(3R,4R,6R)-6-isocyanatomethyl-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazineand 1 ml of benzyl alcohol is stirred at 120° C. for 3 h. The reactionmixture is cooled at RT, and purified by flash chromatography (SiO₂ 60F) to afford the title compound as a yellow resin. Rf=0.17(Heptane/EtOAc 1:1); Rt=5.78 (gradient I).

d)6-[(3R,4R,6R)-6-Isocyanatomethyl-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-1-(toluene-4-sulfonyl)-piperidin-3-yloxymethyl]-4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazine

To a solution of 1 mmol of[(2R,4R,5R)-4-[4-((S)-3-methoxy-2-methyl-propoxymethyl)-phenyl]-5-[4-(3-methoxy-propyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-ylmethoxy]-1-(toluene-4-sulfonyl)-piperidin-2-yl]-aceticacid (example 1g) and 2.5 mmol of Et₃N in 10 ml of THF are added 2 mmolof ethyl chloroformate at 0° C. and the mixture is stirred for 1 h at 0°C. Then a solution of 20 mmol of sodium azide in 2 ml of water is addeddropwise and the reaction mixture is stirred at 0° C. for 1 h. Themixture is diluted with 40 ml of EtOAc and washed with of H₂O (3×). Theorganic layer is dried over Na₂SO₄, and evaporated under reducedpressure. The residue is redissolved in 5 ml of toluene and heated to120° C. for 1.5 h. The toluene is evaporated under reduced pressureyielding the crude title compound as a light brown oil. Rt=5.71(gradient I).

1. A compound of the formula:

or its pharmaceutically acceptable salt, in which R is C₂₋₈-alkenyl,C₂₋₈-alkynyl, C₀₋₈-alkyl-carbonyl-optionally N-mono-C₁₋₈-alkylatedamino-C₁₋₈-alkyl, C₃₋₈-cycloalkyl-C₀₋₈-alkyl,C₁₋₈-alkyl-sulfonyl-C₁₋₈-alkyl, optionally N-mono- orN,N-di-C₁₋₈-alkylated carbamoyl-C₁₋₈-alkyl, optionally O—C₁₋₈-alkylatedcarboxyl-C₀₋₈-alkyl, optionally N and/or N′ mono-, di- ortri-C₁₋₈-alkylated ureido-C₁₋₈-alkyl, heterocyclylcarbonyl-C₀₋₈-alkyl orheterocyclyl-C₀₋₈-alkyl; each of said radicals may be substituted,preferably by 1-4 substituents independently selected from C₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy, C₁₋₈-alkyl, cyano, halogen, hydroxyl, oxo,trifluoromethoxy and trifluoromethyl.
 2. The compound according to claim1, which corresponds to general formulae (IA) or (IB)

or a pharmaceutically acceptable salt thereof, where the meaning of thesubstituent R is as indicated for compounds of the formula (I) accordingto claim
 1. 3. The compound according to claim 1 or 2 in which R isC₀₋₈-alkyl-carbonyl-optionally N-mono-C₁₋₈-alkylated amino-C₁₋₈-alkyl,optionally substituted by 1-2 substituents independently selected fromC₁₋₈-alkoxy, C₁₋₈-alkoxy-C₁₋₈-alkoxy, C₁₋₈-alkyl, cyano, halogen,hydroxyl, oxo, trifluoromethoxy and trifluoromethyl or apharmaceutically acceptable salt thereof.
 4. The compound according toclaim 1 or 2 in which R is optionally N-mono- or N,N-di-C₁₋₈-alkylatedcarbamoyl-C₀₋₈-alkyl, optionally substituted by 1-2 substituentsindependently selected from C₁₋₈-alkoxy, C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl, cyano, halogen, hydroxyl, oxo, trifluoromethoxy andtrifluoromethyl; or a pharmaceutically acceptable salt thereof.
 5. Thecompound according to claim 1 or 2 in which R is optionally N and/or N′mono-, di- or tri-C₁₋₈-alkylated ureido-C₁₋₈-alkyl, optionallysubstituted by 1-2 substituents independently selected from C₁₋₈-alkoxy,C₁₋₈-alkoxy-C₁₋₈-alkoxy, C₁₋₈-alkyl, cyano, halogen, hydroxyl, oxo,trifluoromethoxy and trifluoromethyl; or a pharmaceutically acceptablesalt thereof.
 6. The compound according to claim 1 or 2 in which R isheterocyclyl-C₀₋₈-alkyl, optionally substituted by 1-2 substituentsindependently selected from C₁₋₈-alkoxy, C₁₋₈-alkoxy-C₁₋₈-alkoxy,C₁₋₈-alkyl, cyano, halogen, hydroxyl, oxo, trifluoromethoxy andtrifluoromethyl; or a pharmaceutically acceptable salt thereof.
 7. Thecompound according to claim 1 or 2 in which R isC₀₋₈-alkyl-carbonyl-optionally N-mono-C₁₋₈-alkylated amino-C₁₋₈-alkyloptionally N-mono- or N,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₈-alkyl orheterocyclyl-C₀₋₈-alkyl; or a pharmaceutically acceptable salt thereof.8. The compound according to claim 1, 2 or 7 in which R isC₀₋₄-alkyl-carbonyl-optionally N-mono-C₁₋₄-alkylated amino-C₁₋₄-alkyl,optionally N-mono- or N,N-di-C₁₋₈-alkylated carbamoyl-C₀₋₄-alkyl,morpholinyl-C₀₋₄-alkyl, tetrazolyl-C₀₋₄-alkyl or triazolyl-C₀₋₄-alkyl;or a pharmaceutically acceptable salt thereof.
 9. (canceled)
 10. Apharmaceutical composition comprising a compound according to claim 1;or a pharmaceutically acceptable salt thereof; and optionally one ormore agents having cardiovascular activity.
 11. A method in thetreatment, for the delay of progression or for the prevention of highblood pressure (hypertension), heart failure, glaucoma, myocardialinfarction, renal failure, diabetic nephropathy, stroke or restenosescomprising administering a therapeutically effective amount of acompound according to claim 1; or a pharmaceutically acceptable saltthereof; to a subject in need thereof.